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US 2004/0106184 A1Published application
Chromatographic methods for adenovirus purification
AssigneeIntrogen Therapeutics, Inc.
InventorJoseph Senesac
Life of this patent
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Abstract
This invention provides methods for passing adenovirus particle preparations through chromatographic media to provide purified adenovirus particles.
Claims73
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of:
- (a) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby adenovirus particles from said adenovirus preparation are retained on said first chromatographic medium;
- (b) eluting adenovirus particles from said first chromatographic medium to produce an eluate of adenovirus particles;
- (c) subjecting adenovirus particles from said eluate to chromatography on a second chromatographic medium, wherein said second chromatographic medium retains one or more contaminants from said eluate and wherein said second chromatographic medium is not solely a size exclusion medium;
- and
- (d) collecting adenovirus particles from said eluate.
2 more elements
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of:
- (a) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby contaminants from said adenovirus preparation are retained on said first chromatographic medium;
- (b) subjecting adenovirus particles remaining in the eluant to chromatography on a second chromatographic medium whereby adenovirus particles from said eluant are retained on said second chromatographic medium, wherein when said second chromatographic medium is an anion exchange medium, then said first chromatographic medium is a medium other than a sulfonated polysaccharide affinity medium, and (c) eluting adenovirus particles from said second chromatographic medium.
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of:
- (a) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby contaminants from said adenovirus preparation are retained on said first chromatographic medium;
- (b) subjecting adenovirus particles remaining in the eluant to chromatography on a second chromatographic medium whereby further contaminants are retained on said second chromatographic medium;
- and
- (c) collecting the adenovirus particles remaining in the eluant after step (b).
1 more element
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of:
- (a) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby adenovirus particles from said adenovirus preparation are retained on said first chromatographic medium;
- (b) eluting adenovirus particles from said first chromatographic medium to produce a first eluate of adenovirus particles;
- (c) subjecting said first eluate of adenovirus particles to chromatography on a second chromatographic medium, whereby adenovirus particles from said first eluate are retained on said second chromatographic medium, wherein when said first chromatographic medium is an anion exchange medium, said second chromatographic medium is a medium other than immobilized metal affinity medium, a size exclusion medium, anion exchange medium, cation exchange medium or hydrophobic interaction medium;
- (d) eluting adenovirus particles from said second chromatographic medium to produce a second eluate of adenovirus particles;
- and
- (e) collecting adenovirus particle from said second eluate.
3 more elements
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of:
- (a) preparing said adenoviral preparation according to a method comprising the steps of:
- i) growing host cells in cell culture media
- ii) providing nutrients to said host cells by perfusion, fed-batch, bioreactor, or automated roller bottles;
- iii) infecting said cells with an adenovirus;
- and
- iv) lysing said host cells to provide a cell lysate comprising said adenovirus preparation;
- (b) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby adenovirus particles from said adenovirus preparation are retained on said first chromatographic medium;
- (c) eluting adenovirus particles from said first chromatographic medium to produce an eluate of adenovirus particles;
- (d) subjecting adenovirus particles from said eluate to chromatography on a second chromatographic medium, wherein said second chromatographic medium retains one or more contaminants from said eluate and wherein said second chromatographic medium is not solely a size exclusion medium;
- and
- (e) collecting adenovirus particles from said eluate.
8 more elements
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of:
- (a) preparing said adenoviral preparation according to a method comprising the steps of:
- i) growing host cells in cell culture media
- ii) providing nutrients to said host cells by perfusion, fed-batch, bioreactor, or automated roller bottles;
- iii) infecting said cells with an adenovirus;
- and
- iv) lysing said host cells to provide a cell lysate comprising said adenovirus preparation;
- (b) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby contaminants from said adenovirus preparation are retained on said first chromatographic medium;
- (c) subjecting adenovirus particles remaining in the eluant to chromatography on a second chromatographic medium, whereby adenovirus particles from said eluant are retained on said second chromatographic medium, wherein said first chromatographic medium is a medium other than a sulfonated polysaccharide affinity medium when said second chromatographic medium is an anion exchange medium;
- and
- (d) eluting said adenovirus particles from said second chromatographic medium.
7 more elements
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of:
- (a) preparing said adenoviral preparation according to a method comprising the steps of:
- i) growing host cells in cell culture media
- ii) providing nutrients to said host cells by perfusion, fed-batch, bioreactor, or automated roller bottles;
- iii) infecting said cells with an adenovirus;
- and
- iv) lysing said host cells to provide a cell lysate comprising said adenovirus preparation;
- (b) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby contaminants from said adenovirus preparation are retained on said first chromatographic medium;
- (c) subjecting adenovirus particles remaining in the eluant to chromatography on a second chromatographic medium whereby further contaminants are retained on said second chromatographic medium;
- and
- (d) collecting the adenovirus particles remaining in the eluant after step (c).
7 more elements
A method for preparing adenovirus particles from an adenovirus preparation comprising the steps of;
- (a) preparing said adenoviral preparation according to a method comprising the steps of:
- i) growing host cells in cell culture media
- ii) providing nutrients to said host cells by perfusion, fed-batch, bioreactor, or automated roller bottles;
- iii) infecting said cells with an adenovirus;
- and
- iv) lysing said host cells to provide a cell lysate comprising said adenovirus preparation;
- (b) subjecting said adenovirus preparation to chromatography on a first chromatographic medium, whereby adenovirus particles from said adenovirus preparation are retained on said first chromatographic medium;
- (c) eluting adenovirus particles from said first chromatographic medium to produce a first eluate of adenovirus particles;
- (d) subjecting said first eluate of adenovirus particles to chromatography on a second chromatographic medium, whereby adenovirus particles from said first eluate are retained on said second chromatographic medium, wherein when said first chromatographic medium is an anion exchange medium, then said second chromatographic medium is a medium other than an immobilized metal affinity medium, a size exclusion medium, an anion exchange medium, a cation exchange medium or a hydrophobic interaction medium;
- (e) eluting adenovirus particles from said second chromatographic medium to produce a second eluate of adenovirus particles;
- and
- (f) collecting adenovirus particle from said second eluate.
9 more elements
Dependencies are inferred from the claim text.